HR (homologous recombination) is expected to play important roles in the molecular biology and genetics of archaea, but, so far, few functional properties of archaeal HR have been measured in vivo. In the extreme thermoacidophile Sulfolobus acidocaldarius, a conjugational mechanism of DNA transfer enables quantitative analysis of HR between chromosomal markers. Early studies of this system indicated that HR occurred frequently between closely spaced mutations within the pyrE gene, and this result was later supported by various analyses involving defined point mutations and deletions. These properties of intragenic HR suggested a non-reciprocal mechanism in which donor sequences become incorporated into the recipient genome as short segments. Because fragmentation of donor DNA during cell-to-cell transfer could not be excluded from contributing to this result, subsequent analyses have focused on electroporation of selectable donor DNA directly into recipient strains. For example, S. acidocaldarius was found to incorporate synthetic ssDNA (single-stranded DNA) of more than ∼20 nt readily into its genome. With respect to various molecular properties of the ssDNA substrates, the process resembled bacteriophage λRed-mediated ‘recombineering’ in Escherichia coli. Another approach used electroporation of a multiply marked pyrE gene to measure donor sequence tracts transferred to the recipient genome in individual recombination events. Initial results indicate multiple discontinuous tracts in the majority of recombinants, representing a relatively broad distribution of tract lengths. This pattern suggests that properties of the HR process could, in principle, account for many of the apparent peculiarities of intragenic recombination initiated by S. acidocaldarius conjugation.
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February 2009
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Conference Article|
January 20 2009
Homologous recombination in Sulfolobus acidocaldarius: genetic assays and functional properties
Dennis W. Grogan
Dennis W. Grogan
1
1Department of Biological Sciences, University of Cincinnati, 614 Rieveschl Hall, Mail Location 0006, Cincinnati, OH 45221-0006, U.S.A.
1email grogandw@email.uc.edu
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Publisher: Portland Press Ltd
Received:
July 29 2008
Online ISSN: 1470-8752
Print ISSN: 0300-5127
© The Authors Journal compilation © 2009 Biochemical Society
2009
Biochem Soc Trans (2009) 37 (1): 88–91.
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Received:
July 29 2008
Citation
Dennis W. Grogan; Homologous recombination in Sulfolobus acidocaldarius: genetic assays and functional properties. Biochem Soc Trans 1 February 2009; 37 (1): 88–91. doi: https://doi.org/10.1042/BST0370088
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